Volume 19 Part 1 Article 71: Studies on Tricholoma giganteum and Agaricus bisporus Protoplast Fusion Breeding

Volume 19 Part 1 Article 71
Year 2016
Title: Studies on Tricholoma giganteum and Agaricus bisporus Protoplast Fusion Breeding
Author: Meihua Mo, Dawei Liang, Jiangwei Ma

Abstract:
Tricholoma giganteum is one of the nutritious rare edible fungi. However,its mycelia grows slowly. In order to improve the speed of its growth, the protoplast fusion was conducted using T. giganteum and Agaricus bisporus in this paper.

The condition of protoplast fusion of T. giganteum and A. bisporus was optimized by single factors and uniform design method. The results showed that the optimum protoplast fusion condition was: PEG 26.17%, fusion time 24.4 min, fusion temperature 30.8 ℃, Tris 0.016 mol/L, CaCl2 0.014 mol/L, pH 8.5. The highest fusion rate was 3.87×10-4 at this condition.

One zygote (H49) having good character and the genetic different with their parents was obtained. It was identified from its colony character,morphological state,growth rate and ISSR. The colony character,morphological state of the zygote was different from its parents. Its growth rate was faster than T. giganteum and slower than A. bisporus. Its ISSR genetic correlation coefficients and cluster analysis was analyzed. The result showed that the genetic correlation coefficient between H49 and T. giganteum was 0.65 and between H49 and A. bisporus was 0.47. This indicated that the fusion between T. giganteum and A. bisporus was preliminary successful.

The mycelia growth medium of H49 was optimized. The optimum medium of H49 was: sucrose 1%, peptone 1%, yeast extract 0.11%, VB1 10 mg/L, light calcium carbonate 0.11%, magnesium sulfate 0.29%, potassium dihydrogen phosphate 0.11%; The mycelium of H49 grew fastest (0.98 cm/d) in this medium at its suitable pH 6.

The cultivation property of the fusion strain H49 was studied. It indicated that the mycelium of H49 grew fastest in corn kernels medium. The mycelium was thick and germination time was short. While the cultivation medium of H49 was baggasse 60%, cotton seed hull 30%, wheat bran 9%, yeast extract 0.5%, KH2PO4 (fertilizer grade) 0.3%, MgSO40.1%, light calcium carbonate 0.2%. Its fruiting time was shorter than the parent strain T. giganteum.

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